Supplementary Figure 3: Depletion of DISC1, HZF or SYNCRIP in hippocampal neurons.

(a) DISC1 expression in Disc1^−/− mice. Hippocampal neurons were dissociated from hippocampus of wild-type and Disc1^−/− mice. After 2 d in culture, total protein amounts in the cell lysates were measured by BCA assay followed by immunoblotting with DISC1c-Ab. The asterisks indicate the degradation products of mouse DISC1 recombinant protein (GST-mDISC1). (b,c) Specific knockdown of HZF (b) and SYNCRIP (c) expression by RNAi. Hippocampal neurons were transfected with siRNAs against HZF or SYNCRIP. The graphs represent the densitometry analysis of western blots after normalization to β-tubulin expression. The knockdown efficiencies were calculated in three independent experiments (n = 3). *P < 0.05, Mann-Whitney U-test. Bars show mean ± s.e.m. (d) Effects of HZF and SYNCRIP knockdown on the dendritic localization of ITPR1 3ʹ UTR. Hippocampal neurons were transfected with the indicated siRNAs and Venus–ITPR1 3ʹ UTR plasmids at DIV 12. The transfected neurons were fixed at DIV 14 and immunostained with antibodies to Venus (green) and MAP2 (red). The blue bars indicate the distance between the soma and the farthest transported granules containing ITPR1 3ʹ UTR. The magnified images of these granules are shown in the boxed areas. Scale bars, 15 μm.